Brg1调控重组人骨形态发生蛋白2诱导成骨细胞分化

发布时间：2018-05-22 09:03

  本文选题：成骨 + 染色质重塑　； 参考：《暨南大学》2011年硕士论文

【摘要】：目的：基于原代培养的小鼠颅骨细胞,探索Brg1基因在骨形态发生蛋白2诱导成骨细胞分化过程中的调控机制。 方法：采用胶原酶消化法进行小鼠颅骨成骨细胞的原代培养；分别用0,50,200μg/L的重组人骨形态发生蛋白2诱导原代培养的成骨细胞的分化,摸索骨形态发生蛋白2的最佳作用剂量；实时荧光定量PCR和Western blot进行骨形态发生蛋白2对Brg1的作用时间的动力学分析；实时荧光定量PCR和钙钴染色法检测敲除Brgl对骨形态发生蛋白2诱导的成骨分化的影响；构建Dlx5腺病毒重组表达载体,实时荧光定量PCR和钙钴染色法检测Brg1在骨形态发生蛋白2诱导的成骨分化过程中对Dlx5的调控作用。 结果与结论：用自行合成的重组人骨形态发生蛋白2可诱导原代培养小鼠成骨细胞分化,200μg/L剂量有着较好的诱导分化效果；重组人骨形态发生蛋白2可诱导Brg1基因转录水平和翻译水平表达水平上调；敲除Brgl可抑制重组人骨形态发生蛋白2诱导的成骨分化；Brg1能够调控Dlx5的表达水平。说明Brg1通过调控Dlx5的表达水平调控重组人骨形态发生蛋白2诱导的小鼠成骨细胞的分化。
[Abstract]:Objective: To explore the regulatory mechanism of Brg1 gene in osteoblast differentiation induced by bone morphogenetic protein 2 based on primary cultured mouse cranial cells.
Methods: the primary culture of mouse skull osteoblasts was cultured with collagenase digestion; the differentiation of primary cultured osteoblasts was induced by recombinant human bone morphogenetic protein 2 0,50200 micron g/L, and the optimal dosage of bone morphogenetic protein 2 was explored. Real time fluorescence quantitative PCR and Western blot were used for bone morphogenetic protein 2 pairs. Dynamic analysis of the action time of Brg1; real-time fluorescence quantitative PCR and calcium cobalt staining to detect the effect of knockout Brgl on osteogenic differentiation induced by bone morphogenetic protein 2; construct Dlx5 adenovirus recombinant expression vector, real-time fluorescent quantitative PCR and calcium cobalt staining method to detect Brg1 in the osteogenic differentiation induced by bone morphogenetic protein 2 to D The regulatory role of LX5.
Results and conclusion: the self synthesized recombinant human bone morphogenetic protein 2 could induce the differentiation of osteoblasts in the primary culture of mice. The dose of 200 g/L had a better induction of differentiation. Recombinant human bone morphogenetic protein 2 could induce the up-regulation of Brg1 gene transcription level and the level of translation level, and knockout Brgl could inhibit the morphology of recombinant human bone. The osteogenic differentiation induced by protein 2; Brg1 can regulate the expression level of Dlx5. It shows that Brg1 regulates the differentiation of mouse osteoblasts induced by recombinant human bone morphogenetic protein 2 by regulating the expression level of Dlx5.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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